A corrosion casting/scanning electron microscope method that simultaneously demonstrates clear outlines of endothelial cells and three-dimensional vascular organization.

Abstract

This paper describes a method that can definitively demonstrate endothelial cell boundaries on corrosion casts of arteries, veins, and capillaries. After perfusion with silver nitrate, a casting medium was injected into the entire vascular bed. The injected tissues were either exposed to light or immersed in the photographic developer to develop the silver halide, and corroded in a 5% NaOH solution at 60 degrees C overnight. Observations of the casts containing water in a low vacuum scanning electron microscope equipped with a cooling stage clearly showed endothelial cell boundaries on casts of every type of vessel as well as their three-dimensional architecture. The low vacuum scanning electron microscope images of wet casts were almost identical in quality to the back-scattered electron images of dried casts without any coating. Secondary electron images of the dried casts with metal coating clearly showed endothelial cell outlines and nuclear imprints. The secondary electron images at high magnification indicated that silver granules were precipitated in the grooves along endothelial cell boundaries on the casts. Since this method can demonstrate endothelial cell boundaries of every type of vessel in addition to their three-dimensional architecture, it will be a powerful tool for examining endothelial cell morphology and microvascular organization in pathological as well as normal tissues.